CN104073524B - A kind of method that rich carbon microalgae solid acid diastatic fermentation prepares alcohol fuel - Google Patents
A kind of method that rich carbon microalgae solid acid diastatic fermentation prepares alcohol fuel Download PDFInfo
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- CN104073524B CN104073524B CN201410277518.XA CN201410277518A CN104073524B CN 104073524 B CN104073524 B CN 104073524B CN 201410277518 A CN201410277518 A CN 201410277518A CN 104073524 B CN104073524 B CN 104073524B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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Abstract
The present invention provides a kind of method that rich carbon microalgae solid acid diastatic fermentation prepares alcohol fuel.With rich in carbohydrate microalgae as raw material, under conditions of solid acid is as catalyst, by through centrifugal concentrating micro algae culturing liquid with high-temp liquid water method process, then fermentation preparation produce ethanol.This method is simple and environmentally-friendly, and microalgae is easy to get and grows quickly, and microalgae cell inner cellulose hydrogen bond is more weak, is more easy to be degraded to monosaccharide.Utilize microalgae to prepare ethanol, breach tradition and utilize the raw material such as Semen Maydis, Semen Tritici aestivi to prepare the difficult problem that ethanol cost is high.
Description
Technical field
The present invention relates to technical field prepared by bioenergy, particularly relate to one and utilize microalgae diastatic fermentation to be prepared as second
The process of alcohol ate.
Background technology
Regenerated biological energy seems more and more important in current energy field, and bio-ethanol occupies very important
Status, nowadays fossil liquid fuel price rises steadily, and countries in the world are required for bio-ethanol to supplement energy breach, grind
The persons of studying carefully also are devoted to develop the large-scale production process of bio-ethanol.Two maximum bio-ethanols are raw the most in the world
The Chan Guo U.S. and Brazil, be all with cereal crops as raw material.Bio-ethanol all over the world produces great majority with cereal crops
For raw material, need to take a large amount of arable land, and the consumption of grain also can stimulate food price to go up, bring all multi-environment with
Economic problems.And second filial generation bio-ethanol, i.e. cellulosic ethanol are also faced with the obstacle being much difficult to break through.Exploitation is with non-
Grain crop is one topic received much concern of field of renewable energy as the cheap bio-ethanol of raw material.
Microalgae, as unicellular or simple many cells photosynthetic microorganism, is distributed widely in various environment, can be by photosynthetic
Effect is by solar energy, H2CO in O, air2It is converted into carbohydrate, is stored in microalgae biomass.Microalgae
Growth is with CO in air2For primary carbon source, often produce 1kg microalgae biomass, the CO of 1.83kg can be fixed2.For
Improving the growth rate of microalgae, reduce the discharge of greenhouse gases, people generally utilize industrial waste gas (such as flue gas, little Su
Beat factory's tail gas etc.) in CO2As carbon source.Also have researchers to be processed with waste water by microdisk electrode to combine, profit
With in waste water rich in the feature of the element such as nitrogen, phosphorus, while producing microalgae biomass, process waste water.Open from the eighties
Beginning, under " the water biological species plan " of USDOE is supported, utilizing microalgae to produce fuel progressively becomes global grinding
Study carefully focus and continue up so far.In the last few years, utilize microalgae to prepare ethanol, become each research institution of the world, Energy Section
The focus that skill company is all studied.
Microalgae, compared with traditional raw material preparing bio-ethanol, has that resource is more rich, be more easy to the advantages such as acquisition.Microalgae
The features such as nutritious absorption is fast, photosynthetic efficiency is high, growth is rapid.The photosynthetic efficiency of terrestrial plant is generally lower than 0.5%,
But the photosynthetic efficiency of microalgae reaches as high as 10%.Efficient photosynthetic efficiency makes the growth cycle of microalgae cell shorten, its
Biomass doubling time average out to 25 days, and some algae is only 6 hours, it is possible to produce a large amount of at short notice
Microalgae biomass.By manual control condition, both culturing microalgae can carry out, substantially increases economy, show micro-the whole year
Algae has in terms of as fuel ethyl hydrate raw material and has great advantage.
The raw material of microalgae ethanol is the carbohydrate in microalgae biomass, mainly includes starch, cellulose, hemicellulose
Deng.Many algae such as chlorellas, chlamydomonas, scenedesmus, spirulina etc. are containing substantial amounts of cellulose and starch, and some microalgae forms sediment
Powder content can match in excellence or beauty with other ethanol raw material such as Semen Maydis, Semen Tritici aestivi.Additionally compared with other lignocellulosic plants, microalgae is thin
Intracellular lignin and hemicellulose level are lower, and different from the cellulose I β in plant, are fiber in microalgae cell
Element I α, its hydrogen bond is more weak, is more easy to be degraded to monosaccharide.
Summary of the invention
The present invention provides a kind of method that rich carbon microalgae solid acid diastatic fermentation prepares alcohol fuel,
The technical scheme is that: with rich in carbohydrate microalgae as raw material, under conditions of solid acid is as catalyst,
Being processed by high-temp liquid water method by micro algae culturing liquid through centrifugal concentrating, then fermentation preparation produces ethanol.
The inventive method specifically includes following step:
(1) micro algae culturing liquid through centrifugal concentrating is placed in high-temperature liquid state water treatment facilities, adds water and solid acid,
Hydrolyze 1-3 hour at temperature is 100 DEG C 140 DEG C；The ratio of microalgae dry weight and water is 1:5 1:50；Add consolidates
Body acid is 1:10 1:50 with the ratio of microalgae dry weight；
(2) filtering and reclaim solid acid, the microalgae liquid resin absorption after filtering processes, micro-after adsorption treatment
Algae solution adds 0.5g (NH by every liter of microalgae liquid4)2HPO4、0.025g MgSO4·7H2O, 1.0g yeast extract, adds
Na2HPO4-citrate buffer solution, the pH of regulation reaction system is 4.5-5.5；
(3) cellulase 20-40FPU/g substrate enzyme amount, 50 DEG C of pre-water are added to the reaction system of step (2) gained
Solve 12-24h；
(4) by the inoculum concentration of 5% (v/v) by yeast-inoculated to reaction system, 30 DEG C of fermentation 60-120h, are produced
Thing ethanol.
Specifically, the solid acid in described step (1) is biomass carbon-based solid acid carbon-based solid acid catalysts.Described
The synthetic method of carbon-based solid acid catalyst is as follows: use the materials such as sucrose, glucose, starch in 180 DEG C of carbonizations of temperature
Prepare carbon skeleton carrier, then obtain carbon-based solid acid catalyst by sulfonated reaction at a temperature of 100 DEG C.Use carbon back is solid
Body acid catalyst can be avoided acid corrosion and produce the adverse environmental factors such as substantial amounts of waste water, it is possible to big reduction produces required one-tenth
This.
Specifically, the yeast in described step (4), preferably saccharomyces cerevisiae (Saccharomyces cerevisiae), takes
The Y2034 bacterial strain of slant preservation, with inoculating loop picking one ring yeast thalline, is linked into and trains equipped with 50mLYEPD liquid
Supporting in the 150mL triangular flask of base, be positioned in constant-temperature table, regulation temperature is at 30 DEG C, all under the conditions of 150rpm
Shaken cultivation 12-24 hour.Before being inoculated into fermentation medium, seed liquor 8000rpm taking respective volume is centrifuged 5min,
Outwell supernatant, in inoculation bacterium mud to fermentation flask.Yeast starter culture medium YEPD (w/v): peptone 2%, glucose
2%, yeast powder 1%.
Specifically, described rich in carbohydrate microalgae be carbohydrate content account for microalgae dry weight more than 40%,
Content of starch accounts for the Hi CHO content microalgae kind of more than the 20% of microalgae dry weight.Described microalgae can be room
The inside and outside microalgae kind cultivated.
This method is simple and environmentally-friendly, and microalgae is easy to get and grows quickly, and microalgae cell inner cellulose hydrogen bond is more weak, is more easy to be dropped
Solve as monosaccharide.Utilize microalgae to prepare ethanol, breach tradition and utilize the raw material such as Semen Maydis, Semen Tritici aestivi to prepare the difficulty that ethanol cost is high
Topic.
The technical solution adopted in the present invention compared with prior art, the invention has the beneficial effects as follows that with rich starch microalgae be former
Material, uses the technique that high temperature liquid water process, solid acid catalysis and culture propagation prepare ethanol.Microalgae and traditional raw material
Semen Tritici aestivi, Semen Maydis etc. are compared, and in microalgae cell, lignin and hemicellulose level are lower, and are cellulose I in microalgae cell
α, its hydrogen bond is more weak, is more easy to be degraded to monosaccharide.The present invention is that microalgae biomass application in terms of biomass energy carries
Supply new approach, and solid acid recoverable, simple and environmentally-friendly.This technology is microalgae recovery energy research and probe
New research direction.
Detailed description of the invention
The solid acid used in embodiment is biomass carbon-based solid acid catalyst.The synthesis of described carbon-based solid acid catalyst
Method is as follows: uses the materials such as sucrose, glucose, starch to prepare carbon skeleton carrier in 180 DEG C of carbonizations of temperature, then passes through sulphur
Acidification reaction obtains carbon-based solid acid catalyst, sulfonation time 9 hours, and sulfonation temperature is 100 DEG C.
Embodiment 1:
(1) chlorella algae mud 200g (dry weight 40g) culture fluid through centrifugal concentrating is processed as high temperature liquid water
In device, addition water volume is 400ml, adds solid acid 2g；Temperature of reaction kettle is 120 DEG C of hydrolysis 80min.
(2) filtering and reclaim solid acid, the microalgae liquid resin absorption after filtering processes, and removes the bran in microalgae liquid
The materials such as aldehyde；In the microalgae liquid after adsorption treatment, add inorganic salt material, add 0.5g by every liter of microalgae liquid
(NH4)2HPO4、0.025g MgSO4·7H2O, 1.0g yeast extract, adds Na2HPO4-citrate buffer solution, adjusts
The pH of joint reaction system is 4.5-5.5.
(3) amylase, cellulase 40FPU/g, 50 DEG C of prehydrolysis 24h respectively are added to reaction system.
(4) by the inoculum concentration of 5% (v/v) by saccharomyces cerevisiae (Saccharomyces cerevisiae) NRRL Y-2034
Purchased from northern territory research department of United States Department of Agriculture (USDA), the equal inclined-plane of bacterial strain is stored in 4 DEG C of refrigerators, (YPD fluid medium 30
DEG C cultivate 24h activation) be seeded in reaction system, 30 DEG C fermentation 72h.
Final ethanol 7.32g.
Embodiment 2:
(1) by chlorella dry powder 30g culture fluid as in high-temperature liquid state water treatment facilities, add water to 600ml, add
Solid acid 3g.Temperature of reaction kettle is 140 DEG C of hydrolysis 60min.
(2) filtering and reclaim solid acid, the microalgae liquid resin absorption after filtering processes, and removes the bran in microalgae liquid
The materials such as aldehyde；In the microalgae liquid after adsorption treatment, add inorganic salt material, add 0.5g by every liter of microalgae liquid
(NH4)2HPO4、0.025g MgSO4·7H2O, 1.0g yeast extract, adds Na2HPO4-citrate buffer solution, adjusts
The pH of joint reaction system is 4.5-5.5.
(3) it is separately added into amylase, cellulase 30FPU/g substrate enzyme amount, 50 DEG C of prehydrolysis 18h to reaction system.
(4) by the inoculum concentration of 5% (v) by saccharomyces cerevisiae (Saccharomyces cerevisiae) NRRL Y-2034
Purchased from northern territory research department of United States Department of Agriculture (USDA), the equal inclined-plane of bacterial strain is stored in 4 DEG C of refrigerators, (YPD fluid medium 30 DEG C
Cultivate 24h activation) it is seeded in reaction system, 30 DEG C of fermentation fermentation 120h.
Final ethanol 5.16g.
Embodiment 3
(1) by through centrifugal concentrating for micro-plan ball algae dry powder 50g culture fluid as in high-temperature liquid state water treatment facilities,
Add water 500ml, add solid acid 5g.Temperature of reaction kettle is 100 DEG C of hydrolysis 120min；
(2) filtering and reclaim solid acid, the microalgae liquid resin absorption after filtering processes, and removes the bran in microalgae liquid
The materials such as aldehyde；In the microalgae liquid after adsorption treatment, add inorganic salt material, add 0.5g by every liter of microalgae liquid
(NH4)2HPO4、0.025g MgSO4·7H2O, 1.0g yeast extract, adds Na2HPO4-citrate buffer solution, adjusts
The pH of joint reaction system is 4.5-5.5.
(3) it is separately added into amylase, cellulase 20FPU/g substrate enzyme amount, 50 DEG C of prehydrolysis 24h to reaction system.
(4) by the inoculum concentration of 5% (v), saccharomyces cerevisiae (Saccharomyces cerevisiae) NRRL Y-2034 is purchased
From northern territory research department of United States Department of Agriculture (USDA), the equal inclined-plane of bacterial strain is stored in 4 DEG C of refrigerators and grinds purchased from United States Department of Agriculture (USDA) northern territory
Studying carefully room, the equal inclined-plane of bacterial strain is stored in 4 DEG C of refrigerators, and (24h activation cultivated by YPD fluid medium 30 DEG C) is seeded to reaction
In system, 30 DEG C of fermentation fermentation 120h.
Final ethanol 9.21g.
Embodiment 4:
(1) by chlorella dry powder 10g as in high-temperature liquid state water treatment facilities, addition water volume is 500ml, adds
Solid acid 3g；Temperature of reaction kettle is 100 DEG C of hydrolysis 60min.
(2) filtering and reclaim solid acid, the microalgae liquid resin absorption after filtering processes, and removes the bran in microalgae liquid
The materials such as aldehyde；In the microalgae liquid after adsorption treatment, add inorganic salt material, add 0.5g by every liter of microalgae liquid
(NH4)2HPO4、0.025g MgSO4·7H2O, 1.0g yeast extract, adds Na2HPO4-citrate buffer solution, adjusts
The pH of joint reaction system is 4.5-5.5.
(3) amylase, cellulase 30FPU/g substrate enzyme amount, 50 DEG C of prehydrolysis 24h respectively are added to reaction system.
(4) by the inoculum concentration of 5% (v/v), saccharomyces cerevisiae (Saccharomyces cerevisiae) NRRL Y-2034 is purchased
From northern territory research department of United States Department of Agriculture (USDA), the equal inclined-plane of bacterial strain is stored in 4 DEG C of refrigerators, (YPD fluid medium 30 DEG C training
Support 24h activation) it is seeded in reaction system, 30 DEG C of fermentation 60h.
Final ethanol 1.78g.
Embodiment 5:
(1) by the chlorella powder 40g culture fluid through centrifugal concentrating as in high-temperature liquid state water treatment facilities, water is added
Volume is 200ml, adds solid acid 6g；Temperature of reaction kettle is 120 DEG C of hydrolysis 1800min.
(2) filtering and reclaim solid acid, the microalgae liquid resin absorption after filtering processes, and removes the bran in microalgae liquid
The materials such as aldehyde；In the microalgae liquid after adsorption treatment, add inorganic salt material, add 0.5g by every liter of microalgae liquid
(NH4)2HPO4、0.025g MgSO4·7H2O, 1.0g yeast extract, adds Na2HPO4-citrate buffer solution, adjusts
The pH of joint reaction system is 4.5-5.5.
(3) amylase, cellulase 30FPU/g substrate enzyme amount, 50 DEG C of prehydrolysis 24h respectively are added to reaction system.
(4) by the inoculum concentration of 5% (v/v), saccharomyces cerevisiae (Saccharomyces cerevisiae) NRRL Y-2034 is purchased
From northern territory research department of United States Department of Agriculture (USDA), the equal inclined-plane of bacterial strain is stored in 4 DEG C of refrigerators, (YPD fluid medium 30 DEG C training
Support 24h activation) it is seeded in reaction system, 30 DEG C of fermentation 90h.
Final ethanol 7.25g.
Claims (3)
1. the method that rich carbon microalgae solid acid diastatic fermentation prepares alcohol fuel, with micro-rich in carbohydrate
Algae is raw material, under conditions of solid acid is as catalyst, by the high-temp liquid water of the micro algae culturing liquid through centrifugal concentrating
Method processes, and then fermentation preparation produces ethanol；It is characterized in that: specifically include following step:
(1) micro algae culturing liquid through centrifugal concentrating is placed in high-temperature liquid state water treatment facilities, adds water and solid acid,
Hydrolyze 1-3 hour at temperature is 100 DEG C 140 DEG C；The ratio of microalgae dry weight and water is 1:5 1:50；Add consolidates
Body acid is 1:10 1:50 with the ratio of microalgae dry weight；
(2) filtering and reclaim solid acid, the microalgae liquid resin absorption after filtering processes, micro-after adsorption treatment
Algae solution adds 0.5g (NH by every liter of microalgae liquid4)2HPO4、0.025g MgSO4·7H2O, 1.0g yeast extract, adds
Na2HPO4-citrate buffer solution, the pH of regulation reaction system is 4.5-5.5；
(3) cellulase 20-40FPU/g substrate enzyme amount, 50 DEG C of pre-water are added to the reaction system of step (2) gained
Solve 12-24h；
(4) by 5% inoculum concentration by yeast-inoculated to reaction system, 30 DEG C of fermentation 60-120h, obtain product ethanol；
Solid acid in described step (1) is biomass carbon-based solid acid；The synthesis side of described biomass carbon based solid acid
Method is as follows: use sucrose, glucose or starch to prepare carbon skeleton carrier in 180 DEG C of carbonizations of temperature, then at a temperature of 100 DEG C
Carbon-based solid acid catalyst is obtained by sulfonated reaction.
The method that rich carbon microalgae solid acid diastatic fermentation the most according to claim 1 prepares alcohol fuel, its feature
It is: the yeast in described step (4), is just used for inoculating after 24h activation cultivated by YPD fluid medium 30 DEG C.
The method that rich carbon microalgae solid acid diastatic fermentation the most according to claim 1 prepares alcohol fuel, its feature
It is: described is that carbohydrate content accounts for more than the 40% of microalgae dry weight, starch contains rich in carbohydrate microalgae
Amount accounts for the Hi CHO content microalgae kind of more than the 20% of microalgae dry weight.
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| CN104531779A (en) * | 2014-12-19 | 2015-04-22 | 中国科学院广州能源研究所 | Method for preparing fuel ethanol by fermenting solid alkali saccharification carbon-enriched micro-algae |
| CN104946703A (en) * | 2015-07-07 | 2015-09-30 | 中国科学院广州能源研究所 | Method for treating microalgae saccharification by combining high-temperature liquid water and enzymolysis |
| CN109628502B (en) * | 2018-12-28 | 2022-10-04 | 华东师范大学 | Method for preparing bioethanol by using bloom-forming cyanobacteria as raw material |
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| CN101818217A (en) * | 2010-05-06 | 2010-09-01 | 中国科学院广州能源研究所 | Method and device for pretreatment of high-temperature liquid water of cellulosic biomass |
| CN102247871A (en) * | 2011-05-16 | 2011-11-23 | 中国科学院广州能源研究所 | Preparation method of solid acid catalyst used for biomass hydrolysis |
| CN103421850A (en) * | 2012-05-26 | 2013-12-04 | 北京大学深圳研究生院 | Method used for producing bioethanol with Scenedesmusabundans |
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| CN101818217A (en) * | 2010-05-06 | 2010-09-01 | 中国科学院广州能源研究所 | Method and device for pretreatment of high-temperature liquid water of cellulosic biomass |
| CN102247871A (en) * | 2011-05-16 | 2011-11-23 | 中国科学院广州能源研究所 | Preparation method of solid acid catalyst used for biomass hydrolysis |
| CN103421850A (en) * | 2012-05-26 | 2013-12-04 | 北京大学深圳研究生院 | Method used for producing bioethanol with Scenedesmusabundans |
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